The objectives are to investigate the methylation of RNA in transformed cells and in cells undergoing viral infection. Utilizing the unique features of immunoadsorbents (containing antibodies specific for m6A and for m7G), mRNA molecules and oligonucleotides derived therefrom will be isolated and characterized according to their content of m6A and m7G. Because the above immunoadsorbents can quantitatively and exclusively retain mononucleotides and oligonucleotides (10 to 100 nucleotides in length), studies are in progress to characterize the oligonucleotide sequences of mRNA that contain the corresponding antigenic haptens, i.e., m6A and m7G. Messenger RNAs from hepatoma cell lines and from Adenovirus infected KB cells will be isolated via poly(dT) cellulose chromatography. These RNA will be subjected to a variety of hydrolytic procedures to generate oligonucleotides ranging in size from 50 to 500 nucleotides in length (as characterized by gel electrophoresis). Characterization of those oligonucleotides possessing m7G and/or m6A will be undertaken to determine the location of m6A residues relative to each other, to m6A (5'-end cap) and possibly to the AUG codon. This approach will also permit an analyses of nucleotide sequences adjacent to the 5'-end of mRNA and adjacent to internal m6A residues. BIBLIOGRAPHIC REFERENCES: "Antibodies Specific for N6-Methyladenosine and for 7-Methylguanosine" T.W. Munns, M.K. Liszewski and H.F. Sims, Federation Proceedings, Abstract #2619 (1977). "Characterization of Antibodies Specific for N6-Methyladenosine and for 7-Methylguanosine" T.W. Munns, M.K. Liszewski and H.F. Sims, Biochemistry, May 1977, in press.